Methods of treating diabetes

ABSTRACT

Solid forms of anhydrous (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol are disclosed, in addition to methods of their use in the treatment of various diseases and disorders.

This is a continuation of U.S. application no. 12/503,225, filed Jul.15, 2009, which claims priority to U.S. provisional patent applicationno. 61/081,423, filed July 17, 2008, the entireties of which areincorporated herein by reference.

1. FIELD OF THE INVENTION

This invention relates to solid forms of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol,and to methods of their use.

2. BACKGROUND OF THE INVENTION

Different solid forms of the same compound can have substantiallydifferent properties. For example, the amorphous form of a drug mayexhibit different dissolution characteristics and differentbioavailability patterns than its crystalline form(s), properties whichcan affect how the drug must be administered to achieve optimal effect.Amorphous and crystalline forms of a drug may also have differenthandling properties (e.g., flowability, compressibility), dissolutionrates, solubilities and stabilities, all of which can affect themanufacture of dosage forms. Consequently, access to multiple forms of adrug is desirable for a variety of reasons. Moreover, regulatoryauthorities (e.g., the U.S. Food and Drug Administration) may requirethe identification of all solid (e.g., polymorphic) forms of a new drugsubstance before products containing it. A. Goho, Science News166(8):122-123 (2004).

Compounds may exist in one or more crystalline forms, but the existenceand characteristics of those forms cannot be predicted with anycertainty. In addition, no standard procedure exists for the preparationof all possible polymorphic forms of a compound. And even after onepolymorph has been identified, the existence and characteristics ofother forms can only be determined by additional experimentation. Id.

3. SUMMARY OF THE INVENTION

This invention is directed, in part, to amorphous and crystalline solidforms of anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol,which is an inhibitor of sodium glucose co-transporter 2.

One embodiment of the invention encompasses pharmaceutical compositionscomprising the solid forms described herein.

Another embodiment encompasses methods of inhibiting SGLT2 activity, aswell as methods of treating, preventing and managing a variety ofdiseases and disorders, using the solid forms described herein.

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an X-ray powder diffraction (XRPD) pattern of crystallineanhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol Form 1. The diffractogram was obtained using aBruker D8 Advance System (Cu Kα radiation) with a VANTEC-1 detector.

FIG. 2 is a FT-Raman spectrum of crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolForm 1. The spectrum was obtained using a Bruker RFS100 with 1064 nmexcitation.

FIG. 3 is an XRPD pattern of crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolForm 2. The diffractogram was obtained using a Bruker D8 Advance System(Cu Ka radiation) with a VANTEC-1 detector.

FIG. 4 is a FT-Raman spectrum of crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolForm 2. The spectrum was obtained using a Bruker RFS100 with 1064 nmexcitation.

5. DETAILED DESCRIPTION OF THE INVENTION

This invention is directed, in part, to solid (e.g., crystalline) formsof anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol:

The compound is an inhibitor of the sodium glucose co-transporter 2, andmay be useful in the treatment of diabetes and a variety of otherdiseases and conditions. See U.S. patent application Ser. No.11/862,690, filed Sep. 28, 2007.

This invention is also directed to dosage forms comprising solid formsof anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol,and to methods of their use.

5.1. Definitions

Unless otherwise indicated, the terms “manage,” “managing” and“management” encompass preventing the recurrence of the specifieddisease or disorder in a patient who has already suffered from thedisease or disorder, and/or lengthening the time that a patient who hassuffered from the disease or disorder remains in remission. The termsencompass modulating the threshold, development and/or duration of thedisease or disorder, or changing the way that a patient responds to thedisease or disorder.

Unless otherwise indicated, the terms “prevent,” “preventing” and“prevention” contemplate an action that occurs before a patient beginsto suffer from the specified disease or disorder, which inhibits orreduces the severity of the disease or disorder. In other words, theterms encompass prophylaxis.

Unless otherwise indicated, a “prophylactically effective amount” of acompound is an amount sufficient to prevent a disease or condition, orone or more symptoms associated with the disease or condition, or toprevent its recurrence. A prophylactically effective amount of acompound means an amount of therapeutic agent, alone or in combinationwith other agents, which provides a prophylactic benefit in theprevention of the disease or condition. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

Unless otherwise indicated, a “therapeutically effective amount” of acompound is an amount sufficient to provide a therapeutic benefit in thetreatment or management of a disease or condition, or to delay orminimize one or more symptoms associated with the disease or condition.A therapeutically effective amount of a compound means an amount oftherapeutic agent, alone or in combination with other therapies, whichprovides a therapeutic benefit in the treatment or management of thedisease or condition. The term “therapeutically effective amount” canencompass an amount that improves overall therapy, reduces or avoidssymptoms or causes of a disease or condition, or enhances thetherapeutic efficacy of another therapeutic agent.

Unless otherwise indicated, the terms “treat,” “treating” and“treatment” contemplate an action that occurs while a patient issuffering from the specified disease or disorder, which reduces theseverity of the disease or disorder or one or more of its symptoms, orretards or slows the progression of the disease or disorder.

Unless otherwise indicated, the term “include” has the same meaning as“include, but are not limited to,” and the term “includes” has the samemeaning as “includes, but is not limited to.” Similarly, the term “suchas” has the same meaning as the term “such as, but not limited to.”

Unless otherwise indicated, one or more adjectives immediately precedinga series of nouns is to be construed as applying to each of the nouns.For example, the phrase “optionally substituted alky, aryl, orheteroaryl” has the same meaning as “optionally substituted alky,optionally substituted aryl, or optionally substituted heteroaryl.”

It should also be noted that any atom shown in a drawing withunsatisfied valences is assumed to be attached to enough hydrogen atomsto satisfy the valences. In addition, chemical bonds depicted with onesolid line parallel to one dashed line encompass both single and double(e.g., aromatic) bonds, if valences permit. Structures that representcompounds with one or more chiral centers, but which do not indicatestereochemistry (e.g., with bolded or dashed lines), encompasses purestereoisomers and mixtures (e.g., racemic mixtures) thereof. Similarly,names of compounds having one or more chiral centers that do not specifythe stereochemistry of those centers encompass pure stereoisomers andmixtures thereof.

5.2. Solid Forms

This invention is directed to solid forms of anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol:

One embodiment is directed to solid amorphous forms. Another is directedto solid crystalline forms. A particular crystalline form referred toherein as Form 1 has a differential scanning calorimetry (DSC) endothermat about 124° C. In this context, the term “about” means ±5.0° C. In oneembodiment, the form provides an X-ray powder diffraction (XRPD) patternthat contains peaks at one or more of about 4.0, 8.1, 9.8, 14.0 and/or19.3 degrees 2θ. In this context, the term “about” means ±0.3 degrees.As those skilled in the art are well aware, the relative intensities ofpeaks in an XRPD pattern can vary depending on how the sample isprepared and how the data is collected. With this in mind, an example ofan XRPD pattern of this form is provided in FIG. 1.

In one embodiment, the form provides a Raman spectrum with peaks at oneor more of about 3068, 2929, 2888, 2881, 1615, 1603, 1244, 1037, 692and/or 372 cm⁻¹. In this context, the term “about” means ±2 cm⁻¹. Asthose skilled in the art are well aware, the relative intensities ofpeaks in a Raman spectrum can vary depending on how the sample isprepared and how the data is collected. With this in mind, an example ofa FT-Raman spectrum of this form is provided in FIG. 2.

A particular crystalline form referred to herein as Form 2 has adifferential scanning calorimetry (DSC) endotherm at about 134° C. Inthis context, the term “about” means ±5.0° C. In one embodiment, theform provides an XRPD pattern that contains peaks at one or more ofabout 4.4, 4.8, 14.5, 14.7, 15.5, 21.2, 22.1 and/or 23.8 degrees 20. Inthis context, the term “about” means ±0.3 degrees. An example of an XRPDpattern of this form is provided in FIG. 3.

In one embodiment, the form provides a Raman spectrum with peaks at oneor more of about 3061, 2927, 2877, 2864, 1605, 1038, 842 and/or 719cm⁻¹. In this context, the term “about” means ±2 cm⁻¹. An example of aFT-Raman spectrum of this form is provided in FIG. 4.

This invention encompasses compositions comprising different crystallineforms of anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol.The invention also encompasses solids that are mixtures of bothamorphous and crystalline forms of the compound. Certain such solidscomprise crystalline(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolin an amount of at least about 50, 75, 80, 85, 90, 95 or 99 weightpercent.

5.3. Methods of Use

This invention encompasses a method of inhibiting SGLT2 activity, whichcomprises contacting SGLT2 with an effective amount of a compound of theinvention (i.e., a compound disclosed herein). In one embodiment, theprotein is in vivo. In another, it is ex vivo.

The invention also encompasses a method of decreasing blood glucose in apatient (e.g., a mammal, such as a human, dog or cat), which comprisesadministering to the patient an effective amount of a compound of theinvention.

The invention also encompasses a method of increasing the excretion ofglucose in the urine of a patient, which comprises administering to thepatient an effective amount of a compound of the invention.

The invention also encompasses a method of restoring or increasinginsulin sensitivity in a patient, which comprises administering to thepatient an effective amount of a compound of the invention.

The invention also encompasses a method of treating, managing orpreventing a disease or disorder in a patient, which comprisesadministering to the patient a therapeutically or prophylacticallyeffective amount of a compound of the invention. Examples of diseasesand disorders include atherosclerosis, cardiovascular disease, diabetes(Type 1 and 2), hyperglycaemia, hypertension, lipid disorders, obesity,and Syndrome X. A particular disease is type 2 diabetes.

The amount, route of administration and dosing schedule of a compoundmay depend upon factors such as the specific indication to be treated,prevented or managed, and the age, gender and condition of the patient.The roles played by such factors are well known in the art, and may beaccommodated by routine experimentation.

5.4. Pharmaceutical Formulations

This invention encompasses pharmaceutical compositions comprising one ormore compounds of the invention. Certain pharmaceutical compositions aresingle unit dosage forms suitable for oral, mucosal (e.g., nasal,sublingual, vaginal, buccal, or rectal), parenteral (e.g., subcutaneous,intravenous, bolus injection, intramuscular, or intraarterial), ortransdermal administration to a patient. Examples of dosage formsinclude, but are not limited to: tablets; caplets; capsules, such assoft elastic gelatin capsules; cachets; troches; lozenges; dispersions;suppositories; ointments; cataplasms (poultices); pastes; powders;dressings; creams; plasters; solutions; patches; aerosols (e.g., nasalsprays or inhalers); gels;

liquid dosage forms suitable for oral or mucosal administration to apatient, including suspensions (e.g., aqueous or non-aqueous liquidsuspensions, oil-in-water emulsions, or a water-in-oil liquidemulsions), solutions, and elixirs; liquid dosage forms suitable forparenteral administration to a patient; and sterile solids (e.g.,crystalline or amorphous solids) that can be reconstituted to provideliquid dosage forms suitable for parenteral administration to a patient.

The formulation should suit the mode of administration. For example,oral administration requires enteric coatings to protect the compoundsof this invention from degradation within the gastrointestinal tract.Similarly, a formulation may contain ingredients that facilitatedelivery of the active ingredient(s) to the site of action. For example,compounds may be administered in liposomal formulations, in order toprotect them from degradative enzymes, facilitate transport incirculatory system, and effect delivery across cell membranes tointracellular sites.

The composition, shape, and type of a dosage form will vary depending onits use. For example, a dosage form used in the acute treatment of adisease may contain larger amounts of one or more of the activeingredients it comprises than a dosage form used in the chronictreatment of the same disease. Similarly, a parenteral dosage form maycontain smaller amounts of one or more of the active ingredients itcomprises than an oral dosage form used to treat the same disease. Theseand other ways in which specific dosage forms encompassed by thisinvention will vary from one another will be readily apparent to thoseskilled in the art. See, e.g., Remington's Pharmaceutical Sciences, 18thed., Mack Publishing, Easton Pa. (1990).

5.4.1. Oral Dosage Forms

Pharmaceutical compositions of the invention suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. See, e.g.,Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, EastonPa. (1990).

Typical oral dosage forms are prepared by combining the activeingredient(s) in an intimate admixture with at least one excipientaccording to conventional pharmaceutical compounding techniques.Excipients can take a wide variety of forms depending on the form ofpreparation desired for administration.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms. If desired, tablets can becoated by standard aqueous or nonaqueous techniques. Such dosage formscan be prepared by conventional methods of pharmacy. In general,pharmaceutical compositions and dosage forms are prepared by uniformlyand intimately admixing the active ingredients with liquid carriers,finely divided solid carriers, or both, and then shaping the productinto the desired presentation if necessary. Disintegrants may beincorporated in solid dosage forms to facility rapid dissolution.Lubricants may also be incorporated to facilitate the manufacture ofdosage forms (e.g., tablets).

5.4.2. Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are specifically sterileor capable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

5.4.3. Transdermal, Topical and Mucosal Dosage Forms

Transdermal, topical, and mucosal dosage forms include, but are notlimited to, ophthalmic solutions, sprays, aerosols, creams, lotions,ointments, gels, solutions, emulsions, suspensions, or other forms knownto one of skill in the art. See, e.g., Remington's PharmaceuticalSciences, 16th and 18th eds., Mack Publishing, Easton Pa. (1980 & 1990);and Introduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger,Philadelphia (1985). Transdermal dosage forms include “reservoir type”or “matrix type” patches, which can be applied to the skin and worn fora specific period of time to permit the penetration of a desired amountof active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal, topical, and mucosal dosageforms are well known to those skilled in the pharmaceutical arts, anddepend on the particular tissue to which a given pharmaceuticalcomposition or dosage form will be applied.

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers may be used to assist in delivering active ingredients to thetissue.

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates may also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or hydrates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

6. EXAMPLES

Aspects of this invention can be understood from the following examples.

6.1. Synthesis of((3aS,5R,6S,6aS)-6-hydroxy-2.2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)(morpholino)methanone

To a 12L three-necked round bottom flask with mechanical stirrer, rubberseptum with temperature probe and gas bubbler was charged L-(-)-xylose(504.40 g, 3.360 mol), acetone (5L, reagent grade) and anhydrous MgSO4powder (811.23g, 6.740 mol/2.0 equiv).

The suspension was set stirring at ambient and then concentrated H₂SO₄(50 mL, 0.938 mol/0.28 equiv) was added. A slow mild exotherm wasnoticed (temperature rose to 24° C. over about 1 hr) and the reactionwas allowed to stir at ambient overnight. After 16.25 hours, TLCsuggested all L-xylose had been consumed, with the major product beingthe bis-acetonide along with some(3aS,5S,6R,6aS)-5-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol.The reaction mixture was filtered and the collected solids were washedtwice with acetone (500 mL per wash). The stirring yellow filtrate wasneutralized with concentrated NH4OH solution (39 mL) to pH=8.7. Afterstirring for 10 min, the suspended solids were removed by filtration.The filtrate was concentrated to afford crude bis-acetonide intermediateas a yellow oil (725.23 g). The yellow oil was suspended in 2.5 L waterstirring in a 5L three-necked round bottom flask with mechanicalstirrer, rubber septum with temperature probe and gas bubbler. The pHwas adjusted from 9 to 2 with 1N aq. HCl (142 mL) and stirred at roomtemperature for 6 h until GC showed sufficient conversion of thebis-acetonide intermediate to(3aS,5S,6R,6aS)-5-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol.The reaction was neutralized by the addition of 50% w/w aq. K₂HPO₄ untilpH=7. The solvent was then evaporated and ethyl acetate (1.25L) wasadded to give a white suspension which was filtered. The filtrate wasconcentrated in vacuo to afford an orange oil which was dissolved in 1 Lmethyl tert-butyl ether. This solution had KF 0.23 wt % water and wasconcentrated to afford(3aS,5S,6R,6aS)-5-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol as an orange oil (551.23 g, 86% yield, 96.7 area% pure by GC). ¹H NMR (400 MHz, DMSO-d6) δ 1.22 (s, 3H) 1.37 (s, 3H)3.51 (dd, J=11.12, 5.81 Hz, 1H) 3.61 (dd, J=11.12, 5.05 Hz, 1H)3.93-4.00 (m, 1H) 3.96 (s, 1H) 4.36 (d, J=3.79 Hz, 1H) 4.86 (br. s., 2H)5.79 (d, J=3.54 Hz, 1H). ¹³C NMR (101 MHz, DMSO-d6) δ 26.48, 27.02,59.30, 73.88, 81.71, 85.48, 104.69, 110.73.

To a solution of(3aS,5S,6R,6aS)-5-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol(25.0 g, 131 mmol) in acetone (375 mL, 15×) and H₂O (125 mL, 5×) wasadded NaHCO₃ (33.0 g, 3.0 equiv), NaBr (2.8 g, 20 mol %) and TEMPO (0.40g, 2 mol %) at 20° C. The mixture was cooled to 0-5° C. and solidtrichloroisocyanuric acid (TCCA, 30.5 g, 1.0 equiv) was then added inportions. The suspension was stirred at 20° C. for 24 h. Methanol (20mL) was added and the mixture was stirred at 20° C. for 1 h. A whitesuspension was formed at this point. The mixture was filtered, washedwith acetone (50 mL, 2×). The organic solvent was removed under vacuumand the aqueous layer was extracted with EtOAc (300 mL, 12× x3) and thecombined organic layers were concentrated to afford an oily mixture withsome solid residue. Acetone (125 mL, 5×) was added and the mixture wasfiltered. The acetone solution was then concentrated to afford thedesired acid((3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole-5-carboxylicacid) as a yellow solid (21.0 g, 79%). ¹H NMR (methanol-d4), δ 6.00 (d,J=3.2 Hz, 1H), 4.72 d, J=3.2 Hz, 1H), 4.53 (d, J=3.2 Hz, 1H), 4.38 (d,J=3.2 Hz, 1H), 1.44 (s, 3H), 1.32 (s, 3H).

To a solution of(3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole-5-carboxylicacid (5.0 g, 24.5 mmol) in THF (100 mL, 20×) was added TBTU (11.8 g, 1.5equiv), N-methylmorpholine (NMM, 4.1 mL, 1.5 equiv) and the mixture wasstirred at 20° C. for 30 min. Morpholine (3.2 mL, 1.5 equiv) was thenadded, and the reaction mixture was stirred at 20° C. for an additional6 h. The solid was filtered off by filtration and the cake was washedwith THF (10 mL, 2× x2). The organic solution was concentrated undervacuum and the residue was purified by silica gel column chromatography(hexanes:EtOAc, from 1:4 to 4:1) to afford 4.3 g of the desiredmorpholine amide (64%) as a white solid. ¹H NMR (CDCl3), δ 6.02 (d,J=3.2 Hz, 1H), 5.11 (br s, 1H), 4.62 (d, J=3.2 Hz, 1H), 4.58 (d, J=3.2Hz, 1H), 3.9-3.5 (m, 8H), 1.51 (s, 3H), 1.35 (s, 3H).

6.2. Alternative synthesis of((3aS,5R,6S,6aS)-6-hydroxy-2.2-dimethyltetrahydrofuro[2.3-d][1,3]dioxol-5-yl)(morpholino)methanone

A solution of the diol(3aS,5S,6R,6aS)-5-(hydroxymethyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-olin acetonitrile (5.38 kg, 65% w/w, 3.50 kg active, 18.40 mol),acetonitrile (10.5 L) and TEMPO (28.4 g, 1 mol %) were added to asolution of K₂HPO₄ (0.32 kg, 1.84 mol) and KH₂PO₄ (1.25 kg, 9.20 mol) inwater (10.5 L). A solution of NaClO₂ (3.12 kg, 80% w/w, 27.6 mole, 1.50eq) in water (7.0 L) and a solution of K₂HPO₄ (2.89 kg, 0.90 eq) inwater (3.0 L) were prepared with cooling. Bleach (3.0L, approximate 6%household grade) was mixed with the K₂HPO₄ solution. Approximately 20%of the NaClO₂ solution (1.6 L) and bleach/K₂HPO₄ solution (400 mL, ˜1mol %) were added. The remainders of the two solutions were addedsimultaneously. The reaction mixture turned dark red brown and slowexotherm was observed. The addition rate of the NaClO₂ solution wasabout 40 mL/min (3-4 h addition) and the addition rate for thebleach/K₂HPO₄ solution was about 10-12 mL/min (10 hr addition) whilemaintaining the batch at 15-25° C. Additional charges of TEMPO (14.3g,0.5 mol %) were performed every 5-6 hr until the reaction went tocompletion (usually two charges are sufficient). Nitrogen sweep of theheadspace to a scrubber with aqueous was performed to keep thegreen-yellowish gas from accumulating in the vessel. The reactionmixture was cooled to <10° C. and quenched with Na₂SO₃ (1.4 kg, 0.6 eq)in three portions over 1 hr. The reaction mixture was then acidifiedwith H₃PO₄ until pH reached 2.0-2.1 (2.5-2.7 L) at 5-15° C. The layerswere separated and the aqueous layer was extracted with acetonitrile(10.5 L x3). The combined organic layer was concentrated under vacuo(˜100-120 torr) at <35° C. (28-32° C. vapor, 45-50° C. bath) to lowvolume (˜6-7 L) and then flushed with acetonitrile (40 L) until KF ofthe solution reached <1% when diluted to volume of about 12-15 L withacetonitrile. Morpholine (1.61 L, 18.4 mol, 1.0 eq) was added over 4-6 hand the slurry was aged overnight under nitrogen. The mixture was cooledto 0-5° C. and aged for 3 hours then filtered. The filter cake waswashed with acetonitrile (10 L). Drying under flowing nitrogen gave 4.13kg of the morpholine salt of((3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole-5-carboxylicacid as a white solid (92-94% pure based on ¹H NMR with1,4-dimethoxybenzene as the internal standard), 72-75% yield correctedfor purity. ¹H NMR (D20) δ 5.96 (d, J=3.6 Hz, 1H), 4.58 (d, J=3.6 Hz,1H), 4.53 (d, J=3.2 Hz, 1H), 4.30 (d, J=3.2 Hz, 1H), 3.84 (m, 2H), 3.18(m, 2H), 1.40 (s, 1H), 1.25 (s, 1H). ¹³H NMR (D20) δ 174.5, 112.5,104.6, 84.2, 81.7, 75.0, 63.6, 43.1, 25.6, 25.1.

The morpholine salt of((3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxole-5-carboxylicacid (7.85 kg, 26.9 mol), morpholine (2.40 L, 27.5 mol) and boric acid(340 g, 5.49 mol, 0.2 eq) were added to toluene (31 L). The resultingslurry was degassed and heated at reflux with a Dean-Stark trap undernitrogen for 12 h and then cooled to room temperature. The mixture wasfiltered to remove insolubles and the filter cake washed with toluene (5L). The filtrate was concentrated to about 14 L and flushed with toluene(˜80 L) to remove excess morpholine. When final volume reached ˜12 L,heptane (14 L) was added slowly at 60-70° C. The resulting slurry wascooled gradually to room temperature and aged for 3 h. It was thenfiltered and washed with heptane (12 L) and dry under nitrogen gave aslightly pink solid (6.26 kg, 97% pure, 98% yield). m.p.: 136° C. (DSC).¹H NMR (CDCl₃), δ 6.02 (d, J=3.2 Hz, 1H), 5.11 (br s, 1H), 4.62 (d,J=3.2 Hz, 1H), 4.58 (d, J=3.2 Hz, 1H), 3.9-3.5 (m, 8H), 1.51 (s, 3H),1.35 (s, 3H). ¹³C NMR (methanol-d4) δ 26.84, 27.61, 44.24, 47.45, 68.16,77.14, 81.14, 86.80, 106.87, 113.68, 169.05.

6.3. Synthesis of 1-chloro-2-(4-ethoxybenzyl)-4-iodobenzene

A 2L three-necked round bottom flask with mechanical stirrer, rubberseptum with temperature probe and pressure-equalized addition funnelwith gas bubbler was charged with 2-chloro-5-iodobenzoic acid (199.41 g,0.706 mol), dichloromethane (1.2L, KF=0.003 wt % water) and thesuspension was set stirring at ambient temperature. ThenN,N-dimethylformamide (0.6 mL, 1.1 mol %) was added followed by oxalylchloride (63 mL, 0.722 mol, 1.02 equiv) which was added over 11 min. Thereaction was allowed to stir at ambient overnight and became a solution.After 18.75hours, additional oxalyl chloride (6 mL, 0.069 mol, 0.10equiv) was added to consume unreacted starting material. After 2 hours,the reaction mixture was concentrated in vacuo to afford crude2-chloro-5-iodobenzoyl chloride as a pale yellow foam which will becarried forward to the next step.

A jacketed 2L three-necked round bottom flask with mechanical stirrer,rubber septum with temperature probe and pressure-equalized additionfunnel with gas bubbler was charged with aluminum chloride (97.68 g,0.733 mol, 1.04 equiv), dichloromethane (0.65 L, KF=0.003 wt % water)and the suspension was set stirring under nitrogen and was cooled toabout 6° C. Then ethoxybenzene (90 mL, 0.712 mol, 1.01 equiv) was addedover 7 minutes keeping internal temperature below 9° C. The resultingorange solution was diluted with dichloromethane (75mL) and was cooledto −7° C. Then a solution of 2-chloro-5-iodobenzoyl chloride (≦0.706mol) in 350 mL dichloromethane was added over 13 minutes keeping theinternal temperature below +3° C. The reaction mixture was warmedslightly and held at +5° C. for 2 hours. HPLC analysis suggested thereaction was complete and the reaction was quenched into 450 mLpre-cooled (−5° C.) 2N aq. HCl with stirring in a jacketed round bottomflask. This quench was done in portions over 10 min with internaltemperature remaining below 28° C. The quenched biphasic mixture wasstirred at 20° C. for 45 min and the lower organic phase was washed with1N aq. HCl (200 mL), twice with saturated aq. sodium bicarbonate (200 mLper wash), and with saturated aq. sodium chloride (200 mL). The washedextract was concentrated on a rotary evaporator to afford crude(2-chloro-5-iodophenyl) (4-ethoxyphenyl)methanone as an off-white solid(268.93 g, 99.0 area % by HPLC at 220 nm, 1.0 area % regioisomer at 200nm, 98.5% “as-is” yield).

A jacketed 1 L three-necked round bottom flask with mechanical stirrer,rubber septum with temperature probe and gas bubbler was charged withcrude (2-chloro-5-iodophenyl) (4-ethoxyphenyl)methanone (30.13 g, 77.93mmol), acetonitrile (300 mL, KF=0.004 wt % water) and the suspension wasset stirring under nitrogen and was cooled to about 5° C. Thentriethylsilane (28 mL, 175.30 mmol, 2.25 equiv) was added followed byboron trifluoride-diethyletherate (24 mL, 194.46 mmol, 2.50 equiv) whichwas added over about 30 seconds. The reaction was warmed to ambient over30 min and was stirred for 17 hours. The reaction was diluted withmethyl tert-butyl ether (150 mL) followed by saturated aq sodiumbicarbonate (150 mL) which was added over about 1 minutes. Mild gasevolution was noticed and the biphasic solution was stirred at ambientfor 45 minutes. The upper organic phase was washed with saturated aq.sodium bicarbonate (100 mL), and with saturated aq. sodium chloride (50mL). The washed extract was concentrated on a rotary evaporator to aboutone half of its original volume and was diluted with water (70 mL).Further concentration in vacuo at 45° C. was done until white prillsformed which were allowed to cool to ambient while stirring. After about30 minutes at ambient, the suspended solids were isolated by filtration,washed with water (30 mL), and were dried in vacuo at 45° C. After about2.5 hours, this afforded 1-chloro-2-(4-ethoxybenzyl)-4-iodobenzene as aslightly waxy white granular powder (28.28 g, 98.2 area % by HPLC at 220nm, 97.4% “as-is” yield).

6.4. Synthesis of(4-chloro-3-(4-ethoxybenzyl)phenyl)((3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[[2,3-d]1,3]dioxol-5-yl)methanone

To a solution of 1-chloro-2-(4-ethoxybenzyl)-4-iodobenzene (500 mg, 1.34mmol) in THF (5.0 mL) was added i-PrMgCl (2.0M in THF, 1.0 mL, 2.00mmol) at 0-5° C., and the mixture was stirred for 1.5 h at 0-5° C. Asolution of(3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)(morpholino)methanone(146.5 mg, 0.536 mmol) in THF (1.0 mL) was added dropwise at 0-5° C. andthe mixture was kept stirring for 1 h, warmed to 20° C. and stirred at20° C. for 2 hours. The reaction was quenched with saturated aq NH₄Cl,extracted with MTBE, washed with brine. The organic layer wasconcentrated and the residue was purified by silica gel columnchromatography to afford the desired ketone (178 mg, 76%) as a whitesolid. ¹H NMR (CDCl3) δ 7.88 (dd, J=8.4, 2.0 Hz, 1H), 7.82 (d, J=2.0 Hz,1H), 7.50 (d, J=8.4 Hz, 1H), 7.12 (d, J=8.4 Hz, 2H), 6.86 (d, J=8.4 Hz,2H), 6.07 (d, J=3.2 Hz, 1H), 5.21 (d, J=3.2 Hz, 1H), 4.58 (d, J=3.2 Hz,1H), 4.56 (d, J=3.2 Hz, 1H), 4.16 (d, J=7.2 Hz, 2H), 4.03 (q, J=7.2 Hz,2H), 1.54 (s, 3H), 1.42 (t, J=7.2 Hz, 3H), 1.37 (s, 3H).

6.5. Alternative synthesis of (4-chloro-3-(4-ethmbenzyl)phenyl)((3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)methanone

To a 20 L reactor equipped with a mechanical stirrer, a temperaturecontroller and a nitrogen inlet was charged with the iodide (3.00 kg,8.05 mol) and THF (8 L, 4× to the morpholinoamide) at room temperatureand cooled to −5° C. To the above solution was added dropwise a solutionof i-PrMgCl in THF (Aldrich 2 M, 4.39 L, 8.82 mol) at −5° C. over 3hours. This Grignard solution was used in the ketone formation below.

To a 50 L reactor equipped with a mechanical stirrer, a temperaturecontroller, and a nitrogen inlet was charged the morpholinoamide (HPLCpurity=97 wt %, 2.01 kg, 7.34 mol) and THF (11 L, 5.5×) at roomtemperature and stirred for 45 minutes at room temperature and for 15minutes at 30° C. The homogeneous solution was then cooled to −25° C. Tothis solution was added a solution of t-BuMgCl in THF (Aldrich 1 M, 7.32L, 7.91 mol) at −25° C. over 3 hours. Then the above Grignard solutionwas added to this solution at −20 over 41 minutes. The resultingsolution was further stirred at −20° C. before quench. The reactionmixture was added to 10 wt % aqueous NH₄Cl (10 L, 5×) at 0° C. withvigorous stirring, and stirred for 30 minutes at 0° C. To this mixturewas added slowly 6 N HCl (4 L, 2×) at 0° C. to obtain a clear solutionand stirred for 30 minutes at 10° C. After phase split, the organiclayer was washed with 25 wt % aq NaCl (5 L, 2.5×). Then the organiclayer was concentrated to a 3× solution under the conditions (200 mbar,bath temp 50° C.). EtOAc (24 L, 12×) was added, and evaporated to a 3×solution under the conditions (150 mbar, bath temp 50° C.). Afterremoved solids by a polish filtration, EtOAc (4 L, 2×) was added andconcentrated to dryness (150 mbar, bath temp 50° C.). The wet cake wasthen transferred to a 50 L reactor equipped with a mechanical stirrer, atemperature controller and a nitrogen inlet. After EtOAc was added, thesuspension was heated at 70° C. to obtain a 2.5× homogeneous solution.To the resulting homogeneous solution was added slowly heptane (5 L,2.5×) at the same temperature. A homogeneous solution was seeded andheptane (15 L, 7.5×) was added slowly to a little cloudy solution at 70°C. After stirred for 0.5 h at 70° C., the suspension was slowly cooledto 60° C. and stirred for 1 h at 60° C. The suspension was then slowlycool to room temperature and stirred for 14 h at the same temperature.The crystals were collected and washed with heptane (8 L, 4×), driedunder vacuum at 45° C. to give the desired ketone as fluffy solids (2.57kg, 100 wt % by HPLC, purity-adjusted yield: 81%).

6.6. Synthesis of(2S,3S,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triyltriacetate

To a solution of the ketone(4-chloro-3-(4-ethoxybenzyl)phenyl)-((3aS,5R,6S,6aS)-6-hydroxy-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-5-yl)methanone(114.7 g, 0.265 mol) in MeOH (2 L, 17×) was added CeCl₃.7H₂O (118.5 g,1.2 equiv) and the mixture was stirred at 20° C. until all solids weredissolved. The mixture was then cooled to −78° C. and NaBH₄ (12.03 g,1.2 equiv) was added in portions so that the temperature of the reactiondid not exceed −70° C. The mixture was stirred at −78° C. for 1 hour,slowly warmed to 0° C. and quenched with saturated aq NH₄Cl (550 mL,5×). The mixture was concentrated under vacuum to remove MeOH and thenextracted with EtOAc (1.1L, 10× x2) and washed with brine (550 mL, 5×).The combined organics were concentrated under vacuum to afford thedesired alcohol as a colorless oil (crude, 115 g). To this colorless oilwas added AcOH (650 mL) and H₂O (450 mL) and the mixture was heated to100° C. and stirred for 15 hours. The mixture was then cooled to roomtemperature (20° C.) and concentrated under vacuum to give a yellow oil(crude, ˜118 g). To this crude oil was added pyridine (500 mL) and themixture was cooled to 0° C. Then, Ac20 (195 mL, ˜8.0 equiv) was addedand the mixture was warmed to 20° C. and stirred at 20° C. for 2 h. Thereaction was quenched with H₂O (500 mL) and diluted with EtOAc (1000mL). The organic layer was separated and concentrated under vacuum toremove EtOAc and pyridine. The residue was diluted with EtOAc (1000 mL)and washed with aq NaHSO₄ (1N, 500 mL, x2) and brine (300 mL). Theorganic layer was concentrated to afford the desired tetraacetateintermediate as a yellow foam (˜133g).

To a solution of tetraacetate (133 g, 0.237 mol assuming pure) andthiourea (36.1, 2.0 equiv) in dioxane (530 mL, 4×) was addedtrimethylsilyl trifluoromethanesulfonate (TMSOTf) (64.5 mL, 1.5 equiv)and the reaction mixture was heated to 80° C. for 3.5 hours. The mixturewas cooled to 20° C. and Mel (37 mL, 2.5 equiv) andN,N-diisopropylethylamine (DiPEA) (207 mL, 5.0 equiv) was added and themixture was stirred at 20° C. for 3 h. The mixture was then diluted withmethyl tertiary-butyl ether (MTBE) (1.3 L, 10×) and washed with H₂O (650mL, 5× x2). The organic layer was separated and concentrated undervacuum to give a yellow solid. To this yellow solid was added MeOH (650mL, 5×) and the mixture was reslurried at 60° C. for 2 h and then cooledto 0° C. and stirred at 0° C. for 1 hour. The mixture was filtered andthe cake was washed with MeOH (0° C., 70 mL, x3). The cake was driedunder vacuum at 45° C. overnight to afford the desired triacetate(2S,3S,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triyltriacetate(88 g, 60% over 4 steps) as a pale yellow solid. ¹H NMR (CDCl₃) δ 7.37(d, J=8.0 Hz, 1H), 7.20 (dd, J=8.0, 2.0 Hz, 1H), 7.07 (m, 2H), 6.85 (m,2H), 5.32 (t, J=9.6 Hz, 1H), 5.20 (t, J=9.6 Hz, 1H), 5.05 (t, J=9.6 Hz,1H), 4.51 (d, J=9.6 Hz, 1H), 4.38 (d, J=9.6 Hz, 1h), 4.04 (m, 2H), 2.17(s, 3H), 2.11 (s, 3H), 2.02 (s, 3H), 1.73 (s, 3H), 1.42 (t, J=7.2 Hz,3H).

6.7. Alternative synthesis of(2S,3S,4R,5S,6R)-2-(4-chloro-3-(4-ethmbenzyl)phenyl)-6-(methylthio)tetrahydro-2H-Dyran-3,4,5-triyltriacetate

To a 50 L reactor under nitrogen atmosphere, 40 L MeOH was charged,followed with the ketone (2.50 kg, 5.78 mol) and CeCl₃.7H₂O (2.16 kg,1.0 equiv). Methanol (7.5 L) was added as rinse (totally 47.5 L, 19×). Afreshly prepared solution of NaBH₄ (87.5 g, 0.4 equiv) in aqueous 1 NNaOH (250 mL) was added slowly (35 min) at 15-25° C. The mixture wasthen stirred for 15 min. HPLC analysis of the reaction mixture showedapproximately 90:10 diastereomeric ratio. The reaction was quenched with10 wt % aq NH₄Cl (2.5 L, 1×) and the mixture was concentrated undervacuum to 5×, diluted with water (10 L, 4×) and MTBE (12.5L, 5×). Themixture was cooled to 10° C. and 6 N aq HCl was added until the pH ofthe mixture reached 2.0. Stirring was continued for 10 minutes and thelayers were separated.

The organic layer was washed with H₂O (5L, 2×). The combined aqueouslayer was extracted with MTBE (12.5 L, 5×). The combined organic layerswere washed with brine (2.5 L, 1×) and concentrated under vacuum to 3×.MeCN (15 L, 6×) was added. The mixture was concentrated again to 10 L(4×) and any solid residue was removed by a polish filtration. The cakewas washed with minimal amount of MeCN.

The organic filtrate was transferred to 50 L reactor, and a pre-prepared20 mol % aqueous H₂SO₄solution (61.8 mL 98% concentrated H₂SO₄ and 5 LH₂O) was added. The mixture was heated to 80° C. for 2 hours and thencooled to 20° C. The reaction was quenched with a solution of saturatedaqueous K₂CO₃ (5 L, 2×) and diluted with MTBE (15 L, 6×). The organiclayer was separated, washed with brine (5 L, 2×) and concentrated undervacuum to 5 L (2×). MeCN (12.5 L, 5×) was added and the mixture wasconcentrated to 7.5 L (3×).

The above MeCN solution of(3S,4R,5R,6S)-6-(4-chloro-3-(4-ethoxybenzyl)phenyl)tetrahydro-2H-pyran-2,3,4,5-tetraolwas cooled to 10° C., added with dimethylaminopyridine (17.53 g, 2.5 mol%), followed by slow addition of acetic anhydride (3.23 L, 6.0 equiv)and triethylamine (5 L, 2×, 6.0 equiv) so that the temperature of themixture was kept below 20° C. The reaction was then warmed to 20° C. andstirred for 1 hour and diluted with MTBE (15 L, 6×). The mixture wasslowly quenched with water (7.5 L, 3×). The organic layer was separatedand washed with saturated aqueous KHCO₃ (5L, 2×), 1 N NaHSO₄ (5 L, 2×),and brine (5 L, 2×) in sequence.

The organic layer was then concentrated under vacuum to 5 L (2×). MeCN(12.5 L, 5×) was added and the solution was concentrated to 7.5 L (3×)(KF=0.08%). Dioxane (12.5 L, 5×) was added and the solution wasconcentrated to 7.50 L (3×) (KF=0.02%). Any residual solid was removedby a polish filtration and the cake was washed with minimal amount ofdioxane (500 mL).

To the above filtrate was added thiourea (880 g, 2.0 equiv) and TMSOTf(1.57 L, 1.5 equiv). The reaction mixture was heated to 80° C. for 3hours (>97% conversion). The mixture was cooled to 20° C. and methyliodide (541 mL, 1.5 equiv) and diethylisopropylamine (3.02 L, 3.0 equiv)were added and the mixture was stirred at 20° C. for 18 hours. An extramethyl iodide charge (90 mL, 0.25 equiv) was added and the mixture wasstirred at 20° C. for 1 hours. The mixture was then diluted with MTBE(25 L, 10×) and washed with water (12.5 L, 5× x2). The organic layer wasseparated and concentrated under vacuum to ˜5 L (2×). MeOH (12.5 L, 5×)was added and the mixture was concentrated to 5× to afford a slurry. Themixture was then heated at 60° C. for 1 hour and cooled to 0° C. andstirred at 0° C. for 1 hour. The mixture was filtered and the cake waswashed with MeOH (0° C., 2.5 L, 1× x2, 1.0 L, 0.4×). The cake was driedunder vacuum at 45° C. overnight to afford the desired triacetate (1.49kg, 47% over 4 steps) as a pale yellow/off-white solid.

6.8. Synthesis of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-Dyran-3,4,5-triol

To a slurry of(2S,3S,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triyltriacetate(90.0 g, 0.164mo1) in MeOH (900 mL, 10×) was added NaOMe in MeOH (25 wt%, 18 mL, 0.2×) at 20° C. and the mixture was stirred at 20° C. for 2hours until all solids disappeared. The mixture was then concentrated to300 mL, added to H₂O (1L) and stirred for 1 hour. The solid was filteredand washed with H2O (100 mL, x3) and the cake was dried under vacuum at45° C. overnight to afford the desired methyl thiolate (67.0g, 95%). 1HNMR (CDCI3) δ 7.38 (d, J=8.4 Hz, 1H), 7.22 (m, 2H), 7.11 (d, J=8.8 Hz,2H), 6.83 (d, J=8.8 Hz, 2H), 4.35 (d, J=9.6 Hz, 1H), 4.15 (d, J =9.6 Hz,1H), 4.10-3.95 (m, 3H), 3.64 (t, J=8.8 Hz, 1H), 3.50 (m, 2H), 3.42 (brs, 1H), 2.95 (br s, 1H), 2.57 (br s, 1H), 2.17 (s, 3H), 1.40 (t, J =7.2Hz, 3H).

6.9. Preparation of Crystalline Anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethmbenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3.4.5-triolForm 1

Under slightly positive nitrogen pressure, to a 50 L reactor was chargedMeOH (12 L) and the triacetate (1.70 Kg, 3.09 mol). Methanol (5L) wasadded as a rinse. The slurry was then added NaOMe in MeOH (25 wt %, 340mL, 0.2×) in 15 minutes at 20° C. and the mixture was stirred at 20° C.for 2 hours until all solids disappeared. To the mixture was addedslowly water (25.5 L, 15×) in 45 minutes with 5 g seeding (DSC 123° C.).Solids crashed out and the mixture was stirred at 20° C. for 1 hour,cooled to 0° C. and stirred for 30 minutes. The solid was filtered andwashed with water (1.7 L, 1×, ×2) and the cake was dried under vacuum at45° C. overnight to afford the title compound (m.p. ˜123° C. by DSCpeak; 1.28 Kg, 97.7% yield).

6.10. Preparation of Crystalline Anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethmbenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolForm 2

Under slightly positive nitrogen pressure, to a 50 L reactor was chargedMEK (2-butanone, 4 L) and(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolForm 1 (1.49 Kg). MEK (3.45 L) was added as a rinse. The mixture washeated to 80° C. and heptane (14.9 L, 10×) was slowly added in 1.5hours. Solids started to crash out and the mixture was charged heptane(14.9 L, 10×) in 6 h. The mixture was stirred at 80° C. for 15 hours.The mixture was cooled to 20° C. in 3 hours and stirred at 20° C. for 1hour. The solids were filtered and the cake was washed with MEK/heptane(2.5:7.5, v/v, 1.49 L, 1× x2), dried under nitrogen for 12 hours andunder vacuum at 50° C. for 24 hours to afford the title compound as awhite solid (m.p. ˜134° C. by DSC peak; 1.48 Kg, 98% recovery).

6.11. Alternative Preparation of Crystalline Anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3.4.5-triolForm 2

To a 250 L reactor was charged the triacetate (10 kg) and methanol (75kg). Sodium methoxide (1.6 kg, 30% solution) was added with 5 kgmethanol rinse. The mixture was stirred at room temperature for at least2 hours or until the reaction was complete. Charcoal (Darco G-60, 1 kg)was added with 5 kg methanol rinse. This mixture was heated at 40° C.for 1 h, cooled to room temperature, and filtered through celite. Thecake was washed with methanol (10 kg). Water (100 kg) was added and themixture was concentrated under vacuum. MTBE (200 kg) and water (50 kg)were added and phases were split. The organic layer was washed withwater (100 kg) and concentrated under vacuum. MEK (100 kg) was added andthe same about of solvent was distilled under vacuum. This MEK additionand distillation was repeated to dry the solution. Enough MEK was addedto produce a solution of(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolin 50 L MEK. This solution was polish filtered and heptane (100 L) wasadded at about 80° C. Form 2 seeds (0.1 kg) were added followed by slowaddition of heptane (100 L) as 80° C. Heating was continued for 8 h moreat 80° C., cooled to 20° C. over at least 3 hours, held at thistemperature for at least 2 hours, filtered, and washed with MEK/heptane.The cake was dried at 50° C. under vacuum to afford the title compoundas a white solid (6.6 kg, 86% yield).

All references (e.g., patents and patent applications) cited above areincorporated herein by reference in their entireties.

What is claimed is:
 1. A method of treating diabetes in a patient, whichcomprises administering to a patient in need thereof a therapeuticallyeffective amount of crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyI)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triol.2. The method of claim 1, wherein the diabetes is type 2 diabetes. 3.The method of claim 1, wherein the crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolhas a DSC endotherm at about 124° C.
 4. The method of claim 1, whereinthe crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolhas an X-ray powder diffraction pattern with peaks at one or more ofabout 4.0, 8.1, 9.8, 14.0 and/or 19.3 degrees 2θ.
 5. The method of claim4, wherein the X-ray powder diffraction pattern contains a peak at about14.0 degrees 2θ.
 6. The method of claim 4, wherein the X-ray powderdiffraction pattern is substantially the same as that shown in FIG. 1.7. The method of claim 1, wherein the crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolhas a Raman spectrum with peaks at one or more of about 3068, 2929,2888, 2881, 1615, 1603, 1244, 1037, 692 and/or 372 cm⁻¹.
 8. The methodof claim 7, wherein the Raman spectrum is substantially the same as thatshown in FIG.
 2. 9. The method of claim 1, wherein the crystallineanhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolhas a DSC endotherm at about 134° C.
 10. The method of claim 1, whereinthe crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyI)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolhas an X-ray powder diffraction pattern with peaks at one or more ofabout 4.4, 4.8, 14.5, 14.7, 15.5, 21.2, 22.1 and/or 23.8 degrees 2θ. 11.The method of claim 10, wherein the X-ray powder diffraction pattern hasa peak at about 4.4 degrees
 20. 12. The method of claim 10, wherein theX-ray powder diffraction pattern is substantially the same as that shownin FIG.
 3. 13. The method of claim 1, wherein the crystalline anhydrous(2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-ethoxybenzyl)phenyl)-6-(methylthio)tetrahydro-2H-pyran-3,4,5-triolhas a Raman spectrum with peaks at one or more of about 3061, 2927,2877, 2864, 1605, 1038, 842 and/or 719 cm⁻¹.
 14. The method of claim 13,wherein the Raman spectrum is substantially the same as that shown inFIG. 4.